Fatty Acid Transfer Across the Myocardial Capillary Wall: No Evidence of a Substantial Role for Cytoplasmic Fatty Acid-binding Protein
It has recently been hypothesized that fatty acid (FA) transfer across the myocardial capillary wall is mediated by cytoplasmic fatty acid-binding protein (FABP). Therefore, we studied the type and content of FABP in endothelial cells from rat heart, using molecular biological, immunochemical, and F...
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Veröffentlicht in: | Journal of molecular and cellular cardiology 1994-12, Vol.26 (12), p.1635-1647 |
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creator | Van Nieuwenhoven, Frans A. Verstijnen, Cees P.H.J. Van Eys, Guillaume J.J.M. Van Breda, Eric De Jong, Yvonne F. Van Der Vusse, Ger J. Glatz, Jan F.C. |
description | It has recently been hypothesized that fatty acid (FA) transfer across the myocardial capillary wall is mediated by cytoplasmic fatty acid-binding protein (FABP). Therefore, we studied the type and content of FABP in endothelial cells from rat heart, using molecular biological, immunochemical, and FA-binding assays. Studies were performed on short term cultured endothelial cells, two established endothelial cell lines and ultrathin cryosections from adult rat heart. Northern blotting analysis of endothelial cell RNA failed to detect either heart-type (H-) FABP or liver-type (L-) FABP mRNA, but the reversed transcription-polymerase chain reaction revealed both H- and L- FABP mRNAs, indicating the presence of minor amounts of these mRNAs. Highly sensitive immunochemical assays (sandwich ELISAs) using specific antibodies raised against rat H- or L-FABP showed the contents of these FABP-types in endothelial cells to be 1-5 ng/mg cytosolic protein, which is more than three orders of magnitude lower than the contents of H-FABP in heart or L-FABP in liver. Immuno-electron microscopy also showed that the concentration of H-FABP in endothelial cells is at least two orders of magnitude lower than that in cardiomyocytes. Finally, cytosolic protein samples from endothelial cells revealed no significant FA-binding activity in the 15-kDa region. We conclude that rat heart endothelial cells contain only minor quantities of cytoplasmic FABP and that, therefore, FA transport over the endothelium is mediated by FABP only to a minor extent. It is postulated that aqueous diffusion of FA through the endothelial cytoplasm most likely accounts for the experimentally observed rates of cardiac FA utilization. |
doi_str_mv | 10.1006/jmcc.1994.1183 |
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Therefore, we studied the type and content of FABP in endothelial cells from rat heart, using molecular biological, immunochemical, and FA-binding assays. Studies were performed on short term cultured endothelial cells, two established endothelial cell lines and ultrathin cryosections from adult rat heart. Northern blotting analysis of endothelial cell RNA failed to detect either heart-type (H-) FABP or liver-type (L-) FABP mRNA, but the reversed transcription-polymerase chain reaction revealed both H- and L- FABP mRNAs, indicating the presence of minor amounts of these mRNAs. Highly sensitive immunochemical assays (sandwich ELISAs) using specific antibodies raised against rat H- or L-FABP showed the contents of these FABP-types in endothelial cells to be 1-5 ng/mg cytosolic protein, which is more than three orders of magnitude lower than the contents of H-FABP in heart or L-FABP in liver. Immuno-electron microscopy also showed that the concentration of H-FABP in endothelial cells is at least two orders of magnitude lower than that in cardiomyocytes. Finally, cytosolic protein samples from endothelial cells revealed no significant FA-binding activity in the 15-kDa region. We conclude that rat heart endothelial cells contain only minor quantities of cytoplasmic FABP and that, therefore, FA transport over the endothelium is mediated by FABP only to a minor extent. It is postulated that aqueous diffusion of FA through the endothelial cytoplasm most likely accounts for the experimentally observed rates of cardiac FA utilization.</description><identifier>ISSN: 0022-2828</identifier><identifier>EISSN: 1095-8584</identifier><identifier>DOI: 10.1006/jmcc.1994.1183</identifier><identifier>PMID: 7731058</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>Animals ; Base Sequence ; Biological Transport - physiology ; Blotting, Northern ; Capillaries - metabolism ; Carrier Proteins - metabolism ; Cells, Cultured ; Coronary Vessels - metabolism ; Cytoplasm - metabolism ; Diffusion ; Endothelial cell ; Endothelium, Vascular - metabolism ; Fatty acid transfer ; Fatty acid-binding protein ; Fatty Acid-Binding Protein 7 ; Fatty Acid-Binding Proteins ; Fatty Acids - metabolism ; Immunohistochemistry ; Male ; Molecular Sequence Data ; Neoplasm Proteins ; Nerve Tissue Proteins ; Polymerase Chain Reaction ; Rats ; Rats, Inbred Lew ; Rats, Wistar</subject><ispartof>Journal of molecular and cellular cardiology, 1994-12, Vol.26 (12), p.1635-1647</ispartof><rights>1994 Academic Press</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c339t-70b88c96c49ca1c96ecc77852e104482ec4255ab5bfaf18c6ba104aad11cbe2f3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0022282884711837$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65534</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7731058$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Van Nieuwenhoven, Frans A.</creatorcontrib><creatorcontrib>Verstijnen, Cees P.H.J.</creatorcontrib><creatorcontrib>Van Eys, Guillaume J.J.M.</creatorcontrib><creatorcontrib>Van Breda, Eric</creatorcontrib><creatorcontrib>De Jong, Yvonne F.</creatorcontrib><creatorcontrib>Van Der Vusse, Ger J.</creatorcontrib><creatorcontrib>Glatz, Jan F.C.</creatorcontrib><title>Fatty Acid Transfer Across the Myocardial Capillary Wall: No Evidence of a Substantial Role for Cytoplasmic Fatty Acid-binding Protein</title><title>Journal of molecular and cellular cardiology</title><addtitle>J Mol Cell Cardiol</addtitle><description>It has recently been hypothesized that fatty acid (FA) transfer across the myocardial capillary wall is mediated by cytoplasmic fatty acid-binding protein (FABP). Therefore, we studied the type and content of FABP in endothelial cells from rat heart, using molecular biological, immunochemical, and FA-binding assays. Studies were performed on short term cultured endothelial cells, two established endothelial cell lines and ultrathin cryosections from adult rat heart. Northern blotting analysis of endothelial cell RNA failed to detect either heart-type (H-) FABP or liver-type (L-) FABP mRNA, but the reversed transcription-polymerase chain reaction revealed both H- and L- FABP mRNAs, indicating the presence of minor amounts of these mRNAs. Highly sensitive immunochemical assays (sandwich ELISAs) using specific antibodies raised against rat H- or L-FABP showed the contents of these FABP-types in endothelial cells to be 1-5 ng/mg cytosolic protein, which is more than three orders of magnitude lower than the contents of H-FABP in heart or L-FABP in liver. Immuno-electron microscopy also showed that the concentration of H-FABP in endothelial cells is at least two orders of magnitude lower than that in cardiomyocytes. Finally, cytosolic protein samples from endothelial cells revealed no significant FA-binding activity in the 15-kDa region. We conclude that rat heart endothelial cells contain only minor quantities of cytoplasmic FABP and that, therefore, FA transport over the endothelium is mediated by FABP only to a minor extent. It is postulated that aqueous diffusion of FA through the endothelial cytoplasm most likely accounts for the experimentally observed rates of cardiac FA utilization.</description><subject>Animals</subject><subject>Base Sequence</subject><subject>Biological Transport - physiology</subject><subject>Blotting, Northern</subject><subject>Capillaries - metabolism</subject><subject>Carrier Proteins - metabolism</subject><subject>Cells, Cultured</subject><subject>Coronary Vessels - metabolism</subject><subject>Cytoplasm - metabolism</subject><subject>Diffusion</subject><subject>Endothelial cell</subject><subject>Endothelium, Vascular - metabolism</subject><subject>Fatty acid transfer</subject><subject>Fatty acid-binding protein</subject><subject>Fatty Acid-Binding Protein 7</subject><subject>Fatty Acid-Binding Proteins</subject><subject>Fatty Acids - metabolism</subject><subject>Immunohistochemistry</subject><subject>Male</subject><subject>Molecular Sequence Data</subject><subject>Neoplasm Proteins</subject><subject>Nerve Tissue Proteins</subject><subject>Polymerase Chain Reaction</subject><subject>Rats</subject><subject>Rats, Inbred Lew</subject><subject>Rats, Wistar</subject><issn>0022-2828</issn><issn>1095-8584</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kM1P3DAQxS1URLfAlVsln3rLYudjY_eGVtAi8VEVEEdrMplQoyTe2l6k_Qf4u-uwq3Li5LHemzd6P8ZOpJhLIRanzwPiXGpdzqVUxR6bSaGrTFWq_MRmQuR5lqtcfWZfQngWQuiyKA7YQV0XUlRqxl4vIMYNP0Pb8nsPY-jIp593IfD4h_j1xiH41kLPl7CyfQ9-wx-h77_zG8fPX2xLIxJ3HQd-t25ChDFO5t-uJ945z5eb6FY9hMEif7-VNXZs7fjEf3kXyY5HbL-DPtDx7j1kDxfn98uf2dXtj8vl2VWGRaFjVotGKdQLLDWCTAMh1rWqcpKiLFVOWOZVBU3VdNBJhYsGkgDQSokN5V1xyL5tc1fe_V1TiGawASnVGsmtg6nrWldK6WScb41vKDx1ZuXtkMobKcwE3kzgzQTeTODTwtdd8roZqP1v35FOutrqlOq9WPImoJ3YtdYTRtM6-1H0P00yk7g</recordid><startdate>19941201</startdate><enddate>19941201</enddate><creator>Van Nieuwenhoven, Frans A.</creator><creator>Verstijnen, Cees P.H.J.</creator><creator>Van Eys, Guillaume J.J.M.</creator><creator>Van Breda, Eric</creator><creator>De Jong, Yvonne F.</creator><creator>Van Der Vusse, Ger J.</creator><creator>Glatz, Jan F.C.</creator><general>Elsevier Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19941201</creationdate><title>Fatty Acid Transfer Across the Myocardial Capillary Wall: No Evidence of a Substantial Role for Cytoplasmic Fatty Acid-binding Protein</title><author>Van Nieuwenhoven, Frans A. ; Verstijnen, Cees P.H.J. ; Van Eys, Guillaume J.J.M. ; Van Breda, Eric ; De Jong, Yvonne F. ; Van Der Vusse, Ger J. ; Glatz, Jan F.C.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c339t-70b88c96c49ca1c96ecc77852e104482ec4255ab5bfaf18c6ba104aad11cbe2f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>Animals</topic><topic>Base Sequence</topic><topic>Biological Transport - physiology</topic><topic>Blotting, Northern</topic><topic>Capillaries - metabolism</topic><topic>Carrier Proteins - metabolism</topic><topic>Cells, Cultured</topic><topic>Coronary Vessels - metabolism</topic><topic>Cytoplasm - metabolism</topic><topic>Diffusion</topic><topic>Endothelial cell</topic><topic>Endothelium, Vascular - metabolism</topic><topic>Fatty acid transfer</topic><topic>Fatty acid-binding protein</topic><topic>Fatty Acid-Binding Protein 7</topic><topic>Fatty Acid-Binding Proteins</topic><topic>Fatty Acids - metabolism</topic><topic>Immunohistochemistry</topic><topic>Male</topic><topic>Molecular Sequence Data</topic><topic>Neoplasm Proteins</topic><topic>Nerve Tissue Proteins</topic><topic>Polymerase Chain Reaction</topic><topic>Rats</topic><topic>Rats, Inbred Lew</topic><topic>Rats, Wistar</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Van Nieuwenhoven, Frans A.</creatorcontrib><creatorcontrib>Verstijnen, Cees P.H.J.</creatorcontrib><creatorcontrib>Van Eys, Guillaume J.J.M.</creatorcontrib><creatorcontrib>Van Breda, Eric</creatorcontrib><creatorcontrib>De Jong, Yvonne F.</creatorcontrib><creatorcontrib>Van Der Vusse, Ger J.</creatorcontrib><creatorcontrib>Glatz, Jan F.C.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of molecular and cellular cardiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Van Nieuwenhoven, Frans A.</au><au>Verstijnen, Cees P.H.J.</au><au>Van Eys, Guillaume J.J.M.</au><au>Van Breda, Eric</au><au>De Jong, Yvonne F.</au><au>Van Der Vusse, Ger J.</au><au>Glatz, Jan F.C.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Fatty Acid Transfer Across the Myocardial Capillary Wall: No Evidence of a Substantial Role for Cytoplasmic Fatty Acid-binding Protein</atitle><jtitle>Journal of molecular and cellular cardiology</jtitle><addtitle>J Mol Cell Cardiol</addtitle><date>1994-12-01</date><risdate>1994</risdate><volume>26</volume><issue>12</issue><spage>1635</spage><epage>1647</epage><pages>1635-1647</pages><issn>0022-2828</issn><eissn>1095-8584</eissn><abstract>It has recently been hypothesized that fatty acid (FA) transfer across the myocardial capillary wall is mediated by cytoplasmic fatty acid-binding protein (FABP). Therefore, we studied the type and content of FABP in endothelial cells from rat heart, using molecular biological, immunochemical, and FA-binding assays. Studies were performed on short term cultured endothelial cells, two established endothelial cell lines and ultrathin cryosections from adult rat heart. Northern blotting analysis of endothelial cell RNA failed to detect either heart-type (H-) FABP or liver-type (L-) FABP mRNA, but the reversed transcription-polymerase chain reaction revealed both H- and L- FABP mRNAs, indicating the presence of minor amounts of these mRNAs. Highly sensitive immunochemical assays (sandwich ELISAs) using specific antibodies raised against rat H- or L-FABP showed the contents of these FABP-types in endothelial cells to be 1-5 ng/mg cytosolic protein, which is more than three orders of magnitude lower than the contents of H-FABP in heart or L-FABP in liver. Immuno-electron microscopy also showed that the concentration of H-FABP in endothelial cells is at least two orders of magnitude lower than that in cardiomyocytes. Finally, cytosolic protein samples from endothelial cells revealed no significant FA-binding activity in the 15-kDa region. We conclude that rat heart endothelial cells contain only minor quantities of cytoplasmic FABP and that, therefore, FA transport over the endothelium is mediated by FABP only to a minor extent. It is postulated that aqueous diffusion of FA through the endothelial cytoplasm most likely accounts for the experimentally observed rates of cardiac FA utilization.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>7731058</pmid><doi>10.1006/jmcc.1994.1183</doi><tpages>13</tpages></addata></record> |
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subjects | Animals Base Sequence Biological Transport - physiology Blotting, Northern Capillaries - metabolism Carrier Proteins - metabolism Cells, Cultured Coronary Vessels - metabolism Cytoplasm - metabolism Diffusion Endothelial cell Endothelium, Vascular - metabolism Fatty acid transfer Fatty acid-binding protein Fatty Acid-Binding Protein 7 Fatty Acid-Binding Proteins Fatty Acids - metabolism Immunohistochemistry Male Molecular Sequence Data Neoplasm Proteins Nerve Tissue Proteins Polymerase Chain Reaction Rats Rats, Inbred Lew Rats, Wistar |
title | Fatty Acid Transfer Across the Myocardial Capillary Wall: No Evidence of a Substantial Role for Cytoplasmic Fatty Acid-binding Protein |
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